Journal: Journal of Cell Science

Article Title: Phosphatidylinositol-3-phosphate-dependent Klp98A recruitment regulates endosomal flux underlying developmental synaptic remodeling via Rab4

doi: 10.1242/jcs.264782

Figure Lengend Snippet: Synaptic remodeling in the VNC during third-instar larval stage. (A) Schematic illustrates different parts of the third-instar larval brain. Created in BioRender by Singh, K., 2025. https://BioRender.com/e3oqwsf . This figure was sublicensed under CC-BY 4.0 terms. (B) 3D volume rendered image of a larval VNC stained with anti-Bruchpilot (red, presynaptic junctions) and Hoechst 33342 (white, cortex). Green highlights a single neuromere hemisegment. Representative image of six repeats. (C) Representative images of A3–A6 region of larval VNC from 72–90 h AEL stained with anti-Bruchpilot (nc82) and presented from the look-up table used, which represents intensity in arbitrary units. (D,E) Mean±s.d. of the volume of the synaptic region (D) and Bruchpilot enrichment (intensity, E) in A3–A6 hemisegments marked by anti-Bruchpilot. (F) Representative images of A3–A6 segments of a larval VNC marked by cha>eGFP from 72–90 h AEL and presented from the look-up table used, which represents intensity in arbitrary units. (G,H) Mean±s.d. of GFP intensities and hemisegment volumes (I) and intensity (H) of cholinergic neuromeres marked by endogenous Bruchpilot–GFP. The pairwise significance of the difference is estimated using one-way ANOVA and Tukey's correction for multiple comparisons. A.U., arbitrary units.

Article Snippet: Antibodies, stains and dyes used were: anti:Brp (nc82, DSHB; 1:200), anti-GFP (ab290, Abcam; 1:1000), anti-Rab4 (ab78790, Abcam; 1:400), anti-mouse-IgG conjugated to Alexa Fluor 488 (A11029, A11008, Invitrogen; 1:400) and Hoechst 33342 (Sigma-Aldrich, 1:100).

Techniques: Staining